i want to express a TF with a relatively long Poly Alanine Stretch in bacteria to do GST/His Pulldown experiments. I cloned and started first expression experiments in BL21 cells. Some of my constructs work fine, but those containing the Poly Alanine Stretch seem not to be expressed. Are there any special bacteria out there to deal with more difficult proteins? Already heard of a "Gold" version of those Bl21 cells, but from the manual i dont really get if they are somehow more "competent" in translating difficult proteins...

Any hints/suggestions?

Greetings Tobi

-Tobimpi-

Tobimpi on Feb 26 2009, 12:16 AM said:

Hi,

i want to express a TF with a relatively long Poly Alanine Stretch in bacteria to do GST/His Pulldown experiments. I cloned and started first expression experiments in BL21 cells. Some of my constructs work fine, but those containing the Poly Alanine Stretch seem not to be expressed. Are there any special bacteria out there to deal with more difficult proteins? Already heard of a "Gold" version of those Bl21 cells, but from the manual i dont really get if they are somehow more "competent" in translating difficult proteins...